Translational activation of 5'-TOP mRNA in pressure overload myocardium

The present study was conducted to determine the magnitude and duration of ribosomal protein translation in response to pressure overload and determine if additional, paracrine events associated with mechanical transduction, such as integrin activation using a bioactive peptide ligand, RGD or endothelin stimulation lead to ribosomal protein translation. Polysome analysis of ventricular tissue samples obtained from an in vivo model of right-ventricular pressure overload (RVPO) showed a significant shift in the proportion of a 5'-terminal oligopyrimidine (5'-TOP) mRNA, rpL32, associated with the polysomal fraction when compared with non-5'-TOP mRNAs, beta-actin and beta-myosin heavy chain (beta-MHC), in the early stages of the hypertrophic response (24-48 h). Furthermore, this increase in polysome-bound rpL32 mRNA was accompanied by the phosphorylation of mammalian target of rapamycin (mTOR), p70 S6 kinase (S6K1), and S6 ribosomal protein. In our in vitro studies, treatment of primary cultures of adult feline cardiomyocytes (cardiocytes) with 100 nM endothelin, 9 mM RGD, 100 nM insulin, or 100 nM TPA activated mTOR via distinct signaling pathways and resulted in an increased proportion of polysome-bound rpL32 mRNA. Pre-treatment of cardiocytes with the mTOR inhibitor rapamycin blocked the agonist-induced rpL32 mRNA mobilization to polysomes. These results show that mechanisms that regulate ribosomal biogenesis in the myocardium are dynamically sensitive to pressure overload. Furthermore, our in vitro studies indicate that distinct pathways are operational during the early course of hypertrophic growth and converge to activate mTOR resulting in the translational activation of 5'-TOP mRNA.